刺梨果多糖对肺腺癌A549 细胞增殖、凋亡的作用及机制研究

目的 探讨刺梨果多糖对肺腺癌A549细胞增殖、凋亡的作用及机制。方法 在0、2、4、6、8、10 g·L^-1刺梨果多糖干预A549细胞24、48、72 h后，采用CCK-8法检测A549细胞增殖情况。在0、2、6、10 g·L^-1刺梨果多糖干预A549细胞24、48 h后，采用细胞划痕实验测定A549细胞的迁移情况；Transwell实验测定A549细胞侵袭情况。在0、2、6、10 g·L^-1刺梨果多糖干预A549细胞48 h后，采用流式细胞术(PI单染法)测定A549细胞周期；流式细胞术(Annexin V-FITC/PI双染法)测定A549细胞的凋亡情况；Real-Time PCR法测定A549细胞的周期、凋亡相关基因表达水平；Western Blot法测定A549细胞的周期、凋亡相关蛋白表达水平。结果 (1)干预24、48、72 h后，与对照组(0 g·L^-1)比较，刺梨果多糖2、4、6、8、10 g·L^-1组的A549细胞生长受到显著抑制，细胞的存活率显著降低(P...

Effect and Mechanism Study of Rose roxburghii Tratt. Fruits Polysaccharide on Proliferation andApoptosis of Human Lung Cancer A549 Cells

To investigate the effect and mechanism of Rose roxburghii Tratt. fruits polysaccharide（RTFP） on proliferation and apoptosis of lung adenocarcinoma A549 cells. Methods After A549 cells were treatedwith 0，2，4，6，8，10 g·L-1 RTFP for 24，48，72 hours，the proliferation of A549 cells was detected by CCK-8method. After A549 cells were treated with 0，2，6 and 10 g·L-1 RTFP for 24 and 48 hours，the migration of A549cells was determined by cell scratch test. Transwell assay was used to determine the invasion of A549 cells. The cell cycle of A549 cells was determined by flow cytometry （PI single staining method） after A549 cells were treatedwith 0， 2， 6 and 10 g · L-1 RTFP for 48 hours. The apoptosis of A549 cells was detected by flow cytometry（Annexin V-FITC / PI double staining） . The expression levels of cell cycle and apoptosis-related genes in A549cells were determined by Real-Time PCR. The expression levels of cell cycle and apoptosis-related proteins in A549cells were determined by Western Blot. Results （1） After 24，48 and 72 hours of intervention，compared with thecontrol group （0 g·L-1），the growth of A549 cells in the RTFP 2，4，6，8 and 10 g·L-1 groups was significantlyinhibited，and the survival rate of cells was significantly decreased （P＜0.05，P＜0.01） . The IC50 value of RTFPintervention for 48 hours was 6.109 g·L-1， and the concentration of 2， 6 and 10 g·L-1 was used as the low-，medium- and high- dose groups of RTFP in subsequent experiments. （2） After 48 hours of intervention，comparedwith the control group，the migration distance of A549 cells in the RTFP 2，6 and 10 g·L-1 groups was significantlyincreased （P＜0.05， P＜0.01）， and the number of cell invasion was significantly decreased （P＜0.05， P＜0.01） . The proportion of cells in G0 /G1 phase and G2 /M phase was significantly increased （P＜0.05，P＜0.01），and the proportion of cells in S phase was significantly decreased （P＜0.01） in the 6 and 10 g·L-1 groups. ThemRNA and protein expressions of G0 /G1 and G2 /M phase-related regulatory factors CDK-4， CDK-6， CyclinD1，CDK-1 and CyclinB1 of A549 cells in the 2，6 and 10 g·L-1 groups of RTFP were significantly increased（P＜0.05，P＜0.01） . The apoptosis rate of A549 cells was significantly increased （P＜0.01） . The mRNA expressions ofapoptosis-related genes Caspase-3， Caspase-8 and Caspase-9 were significantly up-regulated （P＜0.05， P＜0.01） . The protein expressions of pro-apoptotic Bax，Caspase-3，Caspase-8 and Caspase-9 were significantly upregulated（P＜0.05， P＜0.01）， and the protein expressions of anti-apoptotic Bcl-2 was significantly downregulated（P＜0.05，P＜0.01） . Conclusion RTFP could inhibit the proliferation，migration and invasion of A549cells， block the cell cycle in G0 /G1 and G2 /M phases， and induce apoptosis of A549 cells through mitochondrialpathway and death receptor pathway.